LOCUS       sequence-462194-        7625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of N-terminal tagged HA-BACH1 (H. sapiens) in human cell 
            lines..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7625)
  AUTHORS   Cao S, Garcia SF, Shi H, James EI, Kito Y, Shi H, Mao H, Kaisari S, 
            Rona G, Deng S, Goldberg HV, Ponce J, Ueberheide B, Lignitto L, 
            Guttman M, Pagano M, Zheng N
  TITLE     Recognition of BACH1 quaternary structure degrons by two F-box 
            proteins under oxidative stress.
  JOURNAL   Cell. 2024 Oct 30:S0092-8674(24)01188-7. doi: 
            10.1016/j.cell.2024.10.012.
  PUBMED    39504958
REFERENCE   2  (bases 1 to 7625)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2024 
            Oct 30:S0092-8674(24)01188-7. doi: 10.1016/j.cell.2024.10.012."
COMMENT     Sequence Label: pcDNA3.1 HA-BACH1
FEATURES             Location/Qualifiers
     source          1..7625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             904..930
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             904..930
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             910..936
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     promoter        complement(3178..3196)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3179..3196)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3216..3233)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3222..3446
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3492..3920
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3579..3598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3789..3810
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3929..3949)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3934..4263
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4114..4249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4176..4195
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4330..5124
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4384..4403)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4994..5013
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5298..5419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5335..5354)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5389..5408
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5468..5484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5468..5484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5481..5503)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5492..5508
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5516..5522,5523..5540,5541..5546))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5561..5582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5699..5716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5870..6458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5950..5969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6629..7420,7421..7489))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7252..7271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7490..7594)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttaccatg
      901 gcctacccct acgacgtgcc cgactacgcc tccctcggat cctctctgag tgagaactcg
      961 gtttttgcct atgaatcttc tgtgcatagc accaatgttt tactcagcct taatgaccag
     1021 cggaagaaag atgtgctgtg cgatgtcacc atctttgtgg agggacagcg gttccgcgct
     1081 caccggtccg tgctggcggc atgcagcagt tacttccact caagaatcgt aggccaggct
     1141 gatggagagc tgaacattac tcttccagaa gaggtgacag ttaaaggatt tgaaccttta
     1201 attcagtttg cctacactgc taaactgatt ttaagtaaag agaatgtgga tgaagtgtgc
     1261 aaatgtgtgg agtttttaag tgtacataat attgaggaat cctgctttca gtttctgaaa
     1321 tttaagtttt tggactccac tgcagaccag caagaatgcc caagaaaaaa atgcttttca
     1381 tcacactgtc agaaaacaga ccttaaactt tcacttttgg accagaggga tctagaaact
     1441 gatgaagtgg aggaatttct ggaaaataaa aatgttcaga ctcctcagtg taaactccgc
     1501 aggtatcaag gaaatgcaaa agcctcacct cctctacaag acagtgccag tcagacatat
     1561 gagtccatgt gcttagagaa ggatgctgct ctggccttgc cttctttatg ccccaaatac
     1621 agaaaattcc aaaaagcatt tggaactgac agagtccgta ctggggaatc tagtgtcaaa
     1681 gacattcatg cttctgttca gccaaatgaa aggtctgaaa atgaatgcct gggaggagtc
     1741 ccggagtgta gagatttgca ggtgatgtta aaatgtgacg aaagtaaatt agcaatggaa
     1801 cctgaagaaa cgaagaaaga tcctgcttct cagtgcccaa ctgaaaaatc agaagtgact
     1861 cctttccccc acaattcttc catagaccct catggacttt attctttgtc tcttttacac
     1921 acatatgacc aatatggtga cttgaatttt gctggtatgc aaaacacaac agtgttaaca
     1981 gaaaagcctt tgtcaggtac agacgtccaa gaaaaaacat ttggtgaaag tcaggattta
     2041 cctttgaaat ccgacttggg caccagggaa gatagtagtg ttgcatctag tgataggagt
     2101 agtgtggagc gagaagtggc agaacaccta gcaaaaggct tctggagtga catttgcagc
     2161 acggacactc cttgccaaat gcagttatca cctgctgtgg ccaaagatgg ctcagaacag
     2221 atctcacaga aacggtctga gtgtccgtgg ttaggtatca ggattagtga gagcccagaa
     2281 ccaggtcaaa ggactttcac aacattaagt tctgtcaact gcccttttat aagtactctg
     2341 agtactgaag gctgttcaag caatttggaa attggaaacg atgattatgt ttcagaaccc
     2401 cagcaagaac cttgcccata tgcttgtgtc attagcttgg gagacgactc tgagacggac
     2461 accgaaggag acagtgaatc ctgttcagcc agagaacaag aatgtgaggt aaaactgcca
     2521 ttcaatgcac aacggataat ttcactgtct cgaaatgatt ttcagtcctt gttgaaaatg
     2581 cacaagctta ctccagaaca gctggattgt atccatgata ttcgaagaag aagtaaaaac
     2641 agaattgctg cacagcgctg tcgcaagaga aaacttgact gtatacagaa tcttgaatca
     2701 gaaattgaga agctgcaaag tgaaaaggag agcttgttga aggaaagaga tcacattttg
     2761 tcaactctgg gtgagacaaa gcagaaccta actggacttt gccagaaagt ttgtaaagaa
     2821 gcagctctga gtcaagaaca aatacagata ctcgccaagt actcagctgc agattgccca
     2881 ctttcatttt taatttctga aaaagataaa agtactcctg atggtgaact ggcgttacca
     2941 tcaattttca gtttatctga ccggcctcca gcagtgctgc ctccctgtgc cagaggaaac
     3001 agtgagcctg gctacgcgcg agggcaggag tcccagcaga tgtccacagc cacctctgag
     3061 caagctgggc ctgcggaaca gtgtcgtcag agtggtggga tctcagattt ctgtcagcag
     3121 atgactgata aatgtactac tgatgagtaa ctcgagcatg catctagagg gccctattct
     3181 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     3241 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3301 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3361 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3421 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3481 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3541 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3601 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3661 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3721 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3781 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3841 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3901 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3961 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4081 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4141 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4201 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4261 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4321 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4381 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4441 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4501 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4561 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4621 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4681 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4741 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4801 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4861 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4921 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4981 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5041 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5101 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5161 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5221 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5281 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5341 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5401 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5461 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5521 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5581 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5641 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5701 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5761 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5821 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5881 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5941 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6001 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6061 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6121 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6181 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6241 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6301 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6361 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6421 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6481 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6541 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6601 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6661 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6721 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6781 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6841 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6901 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6961 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7021 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7081 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7141 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7201 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7261 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7321 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7381 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7441 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7501 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7561 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7621 acgtc
//
